{"doi":"10.1101/2023.03.31.535160","title":"Rescue of blood coagulation Factor VIII exon-16 mis-splicing by antisense oligonucleotides","abstract":"ABSTRACT The human Factor VIII ( F8 ) protein is essential for the blood coagulation cascade and specific F8 mutations cause the rare bleeding disorder Hemophilia A (HA). Here, we investigated the impact of HA-causing single-nucleotide mutations on F8 pre-mRNA splicing. We found that 14/97 (∼14.4%) coding sequence mutations tested in our study induced exon skipping. Splicing patterns of 4/11 (∼36.4%) F8 exons tested were especially sensitive to the presence of common disease-causing mutations. RNA-chemical probing analyses revealed a three-way junction structure at the 3′ end of intron 15 (TWJ-3-15). TWJ-3-15 sequesters the polypyrimidine tract, a key determinant of 3′ splice site strength. Using exon-16 of the F8 gene as a model, we designed specific antisense oligonucleotides (ASOs) that target TWJ-3-15 and identified three that promote the splicing of F8 exon-16. Interaction of TWJ-3-15 with ASOs increases accessibility of the polypyrimidine tract and inhibits the binding of hnRNPA1-dependent splicing silencing factors. Moreover, ASOs targeting TWJ-3-15 rescue diverse splicing-sensitive HA-causing mutations, most of which are distal to the 3’ splice site being impacted. The TWJ-3-15 structure and its effect on mRNA splicing provide a model for HA etiology in patients harboring specific F8 mutations and provide a framework for precision RNA-based HA therapies.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2023,"id":398926,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9635,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1117629,"name":"Guillermo Chacaltana","orcid":null,"position":1,"is_corresponding":false},{"id":842531,"name":"Martin Gutierrez","orcid":"0000-0003-3156-3159","position":2,"is_corresponding":false},{"id":403974,"name":"Nicholas M. Forino","orcid":"0000-0003-1715-6741","position":3,"is_corresponding":false},{"id":1117630,"name":"Arcelia Gonzalez Jimenez","orcid":null,"position":4,"is_corresponding":false},{"id":1117631,"name":"Hanzhang Tao","orcid":null,"position":5,"is_corresponding":false},{"id":1117632,"name":"H. Phong","orcid":null,"position":6,"is_corresponding":false},{"id":626810,"name":"Catherine Oh","orcid":null,"position":7,"is_corresponding":false},{"id":1117633,"name":"Priyanka Chary","orcid":null,"position":8,"is_corresponding":false},{"id":1117634,"name":"I. Quesada","orcid":null,"position":9,"is_corresponding":false},{"id":937593,"name":"Antonia Hamrick","orcid":null,"position":10,"is_corresponding":false},{"id":623022,"name":"Sophie Lee","orcid":"0000-0002-2049-9756","position":11,"is_corresponding":false},{"id":403975,"name":"Michael D. Stone","orcid":"0000-0002-2060-4501","position":12,"is_corresponding":false},{"id":237926,"name":"Jeremy R. Sanford","orcid":"0000-0002-1131-1627","position":13,"is_corresponding":false},{"id":1117145,"name":"Victor Tse","orcid":"0009-0005-6066-3062","position":0,"is_corresponding":true}],"reference_count":59,"raw_metadata":null,"created_at":"2026-07-19T01:19:51.724387Z","pmid":"37034721","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}